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Results for “Repeatability and Reproducibility”

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PubMed1988

Reproducibility of walking test results in chronic obstructive airways disease.

Thirty six patients with chronic airflow obstruction were studied to examine (1) the reproducibility and order effect of repeated walking tests when performed over consecutive days or consecutive weeks; (2) the correlation between walking distance and spirometric measurements; and (3) the effect of static visual clues on performance. In study 1, where 12 patients performed 12 walks over three consecutive days, five minute walking distance increased by 33% between walks 1 and 12, half of the increase occurring after the first three walks. In study 2, where 24 patients performed 12 walks over four consecutive weeks, five minute walking distance increased by 8.5% between walks 1 and 12. A learning effect was seen over the first nine walks. Static visual clues to performance did not affect the distance walked. Spirometric measurements showed no order effect in either study. Although walking distance correlated significantly with FEV1, forced vital capacity, and peak expiratory flow, these measurements were poor predictors of exercise performance. The learning effects seen on repeated performance of walking tests over short intervals should be considered when an individual's response to treatment is being interpreted. When walking tests are used in clinical trials a placebo group or randomised crossover design is essential.

Aged↗

PubMed1993

Respiratory symptoms questionnaire for asthma epidemiology: validity and reproducibility.

BACKGROUND: There is a need for a new respiratory symptoms questionnaire for use in epidemiological research in asthma. METHOD: A questionnaire was designed following a pilot study in 78 subjects. It contains nine questions on symptoms such as wheeze and difficulty with breathing in defined circumstances such as exercise and sleep. It was completed by 211 adults and validated by comparison with a self reported history of asthma and with bronchial hyperresponsiveness to histamine. Its short term reproducibility was measured by three repeat administrations over two weeks. RESULTS: Subjects with asthma (n = 33), particularly those having had an asthma attack in the last year (n = 23), were more likely to report any symptom and to report a greater number of symptoms than those without asthma. The same relationship was found for bronchial hyperresponsiveness and symptoms. Either two or more, or three or more, symptoms appeared to be good indices of self reported asthma and bronchial hyperresponsiveness, or both, with a high sensitivity (65-91%) and specificity (85-96%). Reproducibility was good, with few subjects changing the number of symptoms reported by more than one symptom and none by more than four symptoms. The results compared favourably with those from questions on phlegm production from the MRC questionnaire and were better than those reported for the MRC wheeze questions. CONCLUSIONS: The questionnaire will be useful for epidemiological research on asthma and could form part of a new standardised questionnaire with wide applications.

Adult↗

PubMed1987

Factors affecting antimicrobial susceptibility of Fusobacterium species.

Fifteen clinical isolates of Fusobacterium species were studied to determine their quality of growth on five agar media, their susceptibility to penicillin, cephalothin, cefoxitin, and cefotaxime, the inoculum effect, and the presence of L forms and beta-lactamase. Wilkins-Chalgren agar supported confluent growth best, but Fusobacterium nucleatum exhibited poor growth on all agar media. Most isolates exhibited poor reproducibility of MIC results with repeated agar dilution testing. However, most isolates were susceptible to all antibiotics at the breakpoint concentrations. No inoculum effect was observed, but preparation of an inoculum at a 0.5 McFarland nephelometric standard produced a lower than expected number of CFU (10(6) CFU) in some isolates. L forms were frequently seen. No beta-lactamase was found. The variability in MICs seen with beta-lactam antibiotics was not found when clindamycin was tested. MIC studies with Fusobacterium spp. may be complicated by poor growth on agar media, poor reproducibility, and small inoculum size.

Cefotaxime↗

PubMed2006

Electrochemical determination of arsenite using a gold nanoparticle modified glassy carbon electrode and flow analysis.

A flow analysis electrochemical system has been developed, characterized, and optimized for the determination of arsenite (As(III)). Sensitivity was significantly improved by the electrochemical deposition of gold nanoparticles on a dual glassy carbon electrode, which was inserted into a cross-flow thin-layer electrochemical cell. The electrochemical system was linear up to 15 ppb with a detection limit of 0.25 ppb. Gold deposition was evident from cyclic voltammetry measurements, whereas atomic force microscopy and scanning electron microscopy revealed the size and distribution of deposited gold nanoparticles. The size and density of the nanoparticles were related to the gold salt concentration, deposition time, and potential as well as the electrode position. The response to arsenite was directly related to the frequency, increment, and amplitude of the square wave voltammetry as well as the deposition time and potential. Estimated reproducibility was +/-1.1% at 95% confidence interval for 40 repeated analyses of 8 ppb arsenite during continuous analysis. The reproducibility was far superior if the electrochemical reduction of arsenite was performed in nitric acid instead of hydrochloric or sulfuric acid. The electrochemical system was applicable for analysis of spiked arsenic in mineral water containing a significant amount of various ion elements.

Arsenites↗

PubMed1992

Reproducibility of 24-h growth hormone profiles in children.

The rate of GH secretion and the pattern of GH peaks were compared in a group of nine prepubertal children during their prepubertal period in repeated 24-h GH profiles. At investigation, the children were 6-13 years old (at first profile 6-11 years old) and of normal height (+/- 2 SD). Two profiles were obtained per child, (with a mean time interval of 1.5 years, range 0.7 to 3.5 years. The calculated GH secretions of the first and second profiles were compared. As a group, no significant differences in secreted amount of GH, when expressed as data from the second profile as a percentage of data from the first profile (93 +/- 8%), number of peaks (98 +/- 7%) or mean peak amplitudes (92 +/- 11%), were obtained. Between the repeated curves of an individual child, maximal difference in secretion, number of peaks and mean peak amplitudes ranged around +/- 30%, with a mean intraindividual cv of 12%. The reproducibility in the peak distribution for all profiles was also analysed. The relative frequencies is a percentage of the GH peak amplitudes and peak widths were virtually identical in the repeated profiles. Reproducibility of the temporal pattern of profiles was analysed using time-series analysis (Fourier analysis) and showed no difference in rhythmicity between the different occasions. In conclusion, a high reproducibility of both GH secretion and GH pattern was found for the whole group of prepubertal children.(ABSTRACT TRUNCATED AT 250 WORDS)

Child↗

PubMed1991

Reliability of impedance cardiography in measuring central haemodynamics.

The purpose of the study described here was to investigate the reliability of impedance cardiography (IC) in measuring cardiac output (CO) and central blood volume. Absolute values and changes in these variables obtained by impedance cardiography and by isotope- or thermodilution techniques were compared. The reproducibility of IC within the same day was compared with that of isotope dilution and the reproducibility in IC from day to day was derived. Finally, the effects of the readings of impedance tracings by different observers were quantified. The results are based on 270 measurements in 37 healthy subjects and in 25 unmedicated patients with ischaemic heart disease. We obtained significant correlations between absolute values (y = 0.68x + 1.48) and changes (y = 1.00x + 0.0003) in CO measured by IC and isotope- or thermodilution. IC significantly overestimated absolute values of CO (P less than 0.001). We found a qualitative but no quantitative correlation between thoracic fluid volume measured by IC and central blood volume measured by isotope dilution. IC was highly reproducible both when studies were repeated within the same day (SD on differences in CO = 0.36 1 min-1 for IC; SD on differences in CO = 0.30 1 min-1 for isotope dilution) and on different days (SD on differences in CO = 0.45 1 min-1). A low intra-observer variability was found (SD on differences in CO = 0.12 1 min-1). We conclude that impedance cardiography is reliable in measuring changes in cardiac output and thus suitable for repeated measurements in studies on the haemodynamic effects of physiological or pharmacological intervention. Impedance cardiography is sufficiently reliable for comparison of absolute values of CO between different groups of patients. We cannot recommend impedance cardiography for quantitative studies of central blood volume.

Blood Volume↗

PubMed2003

[Evaluation of psychometric properties of Japanese version of international prostate symptom score and BPH impact index].

PURPOSE: To evaluate psychometric properties of the Japanese version of International Prostate Symptom Score (IPSS) and BPH Impact Index (BII). METHODS: The translated IPSS and BII questionnaires were administered to 103 patients with benign prostatic hyperplasia and 23 asymptomatic men. In 82 patients the questionnaires were repeated 2 weeks later to examine reproducibility. Further 2 weeks later 21 out of 82 patients responded to the questionnaires asking symptoms during the last week rather than during the last month. To evaluated responsiveness the questionnaires were repeated after treatment in 22 patients. Internal consistency, construct validity and criterion validity were examined by proper statistic methods. RESULTS: Reproducibility was good to excellent with weighted kappa more than 0.62 for all items. It was not significantly affected by age, symptom severity, institution type, or whether the patients were asked symptoms during the last week or during the last month. Internal consistency was also good with Cronbach's alpha more than 0.83. Principle component analysis identified 2 components in IPSS and a single component in BII, with all the items contributing to the first component. Most items had significant correlation with external criteria including maximum flow rate, residual urine volume or prostate volume. The scores of patients were clearly larger than asymptomatic men and obviously reduced after receiving therapies. CONCLUSION: Japanese translations of IPSS and BII were shown to be reliable, valid and one-dimensional instruments in the Japanese patients. They would be equivalent to the original English questionnaires.

Aged↗

PubMed1996

Ambulatory blood pressure reproducibility and application of the method in a healthy Finnish cohort.

The aim of the present study has been to study the reproducibility, validity and normal values of ambulatory blood pressure measurement in a healthy Finnish cohort. The reproducibility of ambulatory blood pressure monitoring was examined in healthy volunteers and normal values were determined in a Finnish cohort of males and women of different ages. In the reproducibility study the recording was repeated at 2-week intervals. In the validity study simultaneous measurements were done manually with a mercury sphygmomanometer and compared with the measurement by the ambulatory blood pressure recording unit, both connected with the cuff by a T-tube; this study included 100 consecutive measurements in a patient group. A relatively small cohort was taken from the normal value study group. In addition to ambulatory blood pressure their left ventricle mass was also determined by echocardiography. The correlation between manual blood pressure measurement and simultaneous measurement by the ambulatory blood pressure unit was 0.98. In the reproducibility study the correlations between the two 24-h measurements 2 weeks apart were also good. Depending on the parameter (daytime, nighttime or 24-h blood pressure mean, systolic or diastolic values) the correlation coefficient varied between 0.81 and 0.91. Thus both systolic and diastolic blood pressures, whether expressed as 24-h averages and daytime or nighttime averages, correlated well between these two recording sessions. The data obtained in the present Finnish cohort were well within the published reference value limits, showing only fairly modest age-dependence appearing at quite an old age. There was no significant correlation between the left ventricle mass and ambulatory blood pressure values in a population having normal blood pressure. The results suggest good intra-individual correlation and reproducibility in ambulatory blood pressure monitoring, suggesting this method to be useful in the monitoring of individual blood pressure levels. The validity of the method is good and the published reference values can be applied with reasonable reliability in different populations. In an adult population with normal blood pressure, no correlation between left ventricle mass and blood pressure values can be found, suggesting this correlation may first appear in cohorts including persons with elevated blood pressure values.

Adolescent↗

PubMed1998

[Reproducibility and wave analysis of multifocal electroretinography].

We studied the reproducibility of multifocal electroretinography by repeated measurements in normal subjects. We also analyzed the amplitude and implicit time of the waves. Nine healthy eyes including myopic cases (-4.1 D on average) of 9 normal males, aged 19 to 42 years, were the objects of study. We used a (Visual Evoked Response Imaging System III (VERIS III), by TOMEY Corp.) for multifocal electroretinography. Each eye was examined 4 times on different days. The stimulus elements consisted of 103 hexagons. The net recording time was 4 minutes. We used the coefficient of variation (standard deviation/average) of 4 measurements of response density as index of reproducibility. Poor fixation and blinking were important factors that affected the reproducibility. In all the 9 eyes, reproducibility of multifocal electroretinography, expressed as average of the coefficient of variation of all regions, was 22% on average. In 6 eyes with good fixation and without blinking, the reproducibility was 15% on average. In 6 eyes with good reproducibility, we analyzed three parameters of the focal response waves: all traces, quadrants and rings, as well as the amplitude and latency of the waves. These data served as controls. Our studies showed that the response density correlated closely with the amplitude and not with the latency in normal subjects.

Adult↗

PubMed2006

The effect of signal averaging on the reproducibility and reliability of measures of T-wave morphology.

Electrocardiographic repolarization abnormalities characterized by T-wave morphology parameters such as the principal component analysis ratio and the relative and the absolute T-wave residuum (TWR(rel) and TWR(abs)) are predictive of cardiovascular and/or all-cause mortality. However, when using a "10-second median beat" for analysis, the reported mean values for TWR(rel) vary widely and parameter reproducibility is somewhat suspect. In repeated electrocardiographic recordings conducted 1 month and 1 year apart on 15 and 27 healthy individuals, respectively, we studied the said T-wave morphology parameters in single complexes and in reduced noise signal averages containing 10 and 200 complexes. Considering all subjects, the mean (+/-SD) TWR(rel) was highest in a single complex (0.0345% +/- 0.0183%), intermediate in the 10-beat signal-averaged complexes (0.0125% +/- 0.0051%), and lowest in the 200-beat signal-averaged complexes (0.0078% +/- 0.0036%) (P < .0001), with the same trend also observed in the TWR(abs) but not in the principal component analysis ratio. Reproducibility as quantified by within-subject variance and reliability as quantified by the intraclass (intrasubject) correlation coefficient also improved as the number of T-wave complexes analyzed increased. We conclude that signal averages consisting of more than 10 complexes (or more than 10 seconds worth of complexes) are required to produce reproducible and reliable values for TWR(rel) and TWR(abs).

Action Potentials↗

PubMed2000

Comparison of cardiac output measured by two automated methods of CO2 rebreathing.

PURPOSE: The aim of the present study was to investigate the reproducibility of the exponential method of CO2 rebreathing with the use of automated curve fitting and to determine whether this method is superior to the equilibrium method in terms of reproducibility and clinical practicability. METHODS: Repeated measurements of cardiac output were performed using the automated equilibrium and exponential methods. These measurements were compared in 12 healthy male subjects at rest and during incremental exercise tests. RESULTS: Estimated cardiac output was not significantly different between duplicate measurements at rest nor at any level of exercise with either method. At rest the exponential method showed a tendency toward larger variability than the equilibrium method. The exponential method produced significantly higher (P < or = 0.001) estimates at rest (averaging up to 9.8 L x min(-1)) compared with the equilibrium method (averaging up to 6.5 L x min(-1)). Reproducibility improved for both methods with increasing workloads, and a second measurement at rest also seemed more reproducible and valid than the first. During exercise, both methods produced comparable values for cardiac output, and highly significant relations between cardiac output and oxygen uptake were observed for both methods (ranging from r2 = 0.79 to r2 = 0.88, P < or = 0.001). The equilibrium method produced unpleasant side effects more frequently (75% vs 21%, P < or = 0.001) compared with the exponential method and lead more subjects to premature interruption of the exercise test because of the rebreathing maneuver (42% vs 17%, P = 0.058). CONCLUSIONS: Automated curve fitting for the exponential method gave reproducible and valid results during submaximal and maximal exercise but not at rest. The equilibrium method on the other hand interfered with exercise. Therefore, the equilibrium method is recommended at rest and at lower levels of exercise and the exponential method at higher intensities.

Adolescent↗

PubMed2006

Myotonic dystrophy expanded CUG repeats disturb the expression and phosphorylation of tau in PC12 cells.

Mental retardation is a main feature of the congenital form of myotonic dystrophy (DM1), however, the molecular mechanisms underlying the central nervous system symptoms of DM1 are poorly understood. We have established a PC12 cell line-based model expressing the DM1 expanded CUG repeats (CTG90 cells) to analyze the effects of this mutation on neuronal functions. Previously, we have reported that CTG90 cells displayed impaired NGF-induced neuronal differentiation. Because disruption of normal expression of the microtubule associated protein tau and neuronal aggregates of hyperphosphorylated tau have been associated with DM1, this study analyzes the behavior of tau in the CTG90 cells. Several alterations of tau were observed in the PC12 cells that express expanded CUG repeats, including a subtle but reproducible reduction in the expression of the tau mRNA splicing isoform containing exon 10, decreased expression of tau and hyperphosphorylation of both tau and high molecular weight tau as well as abnormal nuclear localization of tau phosphorylated at Ser396/404. Interestingly, phosphorylation regulates negatively the activity of tau as microtubule-associated protein. In addition, impaired activity of the Akt/GSK3beta pathway, which phosphorylates tau, was also identified in the CTG90 cells. Besides tau phosphorylation, the Akt/GSK3beta signaling pathway regulates other key processes of PC12 cells, such as apoptosis and neuronal differentiation. Our results indicate that defective neuronal differentiation exhibited by the PC12 cells expressing expanded CUG repeats could be the result of combinatory effects derived from the altered behavior of tau and the impaired activation of the Akt/GSK3beta signaling pathway.

Animals↗

PubMed2002

Meal and oral glucose tests for assessment of beta -cell function: modeling analysis in normal subjects.

We investigated beta-cell function and its relationship to insulin sensitivity in 17 normal volunteers. For insulin secretion (derived by C-peptide deconvolution), a mathematical model was applied to 24-h triple-meal tests (MT) as well as oral glucose tolerance tests (OGTT); insulin sensitivity was assessed by the euglycemic insulin clamp technique. The beta-cell model featured a glucose concentration-insulin secretion dose response (characterized by secretion at 5 mM glucose and slope), a secretion component proportional to the glucose concentration derivative, and a time-dependent potentiation factor (modulating the dose response and accounting for effects of sustained hyperglycemia and incretins). The beta-cell dose-response functions estimated from the whole 24-h MT, the first 2 h of the MT, and the OGTT differed systematically, because a different potentiation factor was involved. In fact, potentiation was higher than average during meals (1.6 +/- 0.1-fold during the first meal) and had a different time course in the MT and OGTT. However, if potentiation was accounted for, the 24- and 2-h MT and the OGTT yielded similar dose responses, and most beta-cell function parameters were intercorrelated (r = 0.50-0.86, P < or = 0.05). The potentiation factor was found to be related to plasma glucose-dependent insulin-releasing polypeptide concentrations (r = 0.49, P < 0.0001). Among beta-cell function parameters, only insulin secretion at 5 mM glucose from MT correlated inversely with insulin sensitivity (24-h MT: r = -0.74, P < 0.001; 2-h MT: r = -0.52, P < 0.05), whereas the dose-response slope and the OGTT parameters did not. In nine other subjects, reproducibility of model parameters was evaluated from repeated MTs. Coefficients of variation were generally approximately 20%, but the derivative component was less reproducible. We conclude that our model for the multiple MT yields useful information on beta-cell function, particularly with regard to the role of potentiation. With cautious interpretation, a 2-h MT or a standard OGTT can be used as surrogates of 24-h tests in assessing spontaneous beta-cell function.

Adult↗

PubMed2004

Dose-response effect of a single administration of oral hexyl-insulin monoconjugate 2 in healthy nondiabetic subjects.

OBJECTIVE: 1) To evaluate the effect of a single oral dose of hexyl-insulin monoconjugate 2 (HIM2) on the rate of whole-body glucose disposal (Rd) and endogenous glucose production (EGP) in healthy nondiabetic subjects, 2) to examine the reproducibility of HIM2 on glucose metabolism, and 3) to compare the results obtained with HIM2 with those using a bioequivalent dose of subcutaneous lispro insulin. RESEARCH DESIGN AND METHODS: Six healthy subjects ([means +/- SE] aged 31 +/- 5 years and BMI 23.1 +/- 3.9 kg/m2) participated in four studies performed in random order on separate days. Subjects ingested a single dose of HIM2 (0.125, 0.5, and 0.75 mg/kg) or received subcutaneous lispro insulin (0.1 units/kg). Studies were performed with [3-3H]glucose, and plasma glucose concentration was maintained at basal levels for 4 h with the euglycemic clamp technique. After 6 weeks, subjects participated in two repeat studies to examine the reproducibility of HIM2 (0.5 mg/kg) and lispro insulin (0.1 units/kg). RESULTS: Fasting plasma insulin (7 muU/ml) increased to a maximum of 102, 321, and 561 muU/ml at 60 min after all three HIM2 doses (0.125, 0.5, and 0.75 mg/kg, respectively). A dose-related decrease in basal EGP was observed as the HIM2 dosage was increased from 0 to 0.125 to 0.5 mg/kg (P <0.05 vs. each preceding dose). Suppression of EGP was similar with the 0.5- and 0.75-mg/kg HIM2 doses. A dose-related stimulation of basal Rd was observed as the HIM2 dosage was increased from 0 to 0.125 to 0.5 (P <0.05 vs. each preceding dose) to 0.75 mg/kg (P <0.10 vs. preceding dose). Rd (0-240 min) was increased by 0.5 mg/kg oral HIM2 to a value similar to 0.1 units/kg lispro insulin. The 0.125-mg/kg HIM2 dose reduced EGP (0-240 min) to a value that was similar to 0.1 units/kg lispro insulin. The variability in the effect of HIM2 and lispro on Rd (25 +/- 7 vs. 27 +/- 1%, respectively) and on suppression of EGP (19 +/- 1 vs. 19 +/- 0.7%, respectively) was similar. CONCLUSIONS: Oral HIM2 suppresses EGP and increases tissue Rd in a dose-dependent manner. The effects of HIM2 on EGP and Rd persisted at 240 min, even though plasma insulin concentration had returned to basal levels. Oral HIM2 may provide an effective and reproducible means of controlling postprandial plasma glucose excursions in diabetic patients.

Administration, Oral↗

PubMed2002

An international intercomparison exercise for the determination of purified microcystin-LR and microcystins in cyanobacterial field material.

The comparability of current microcystin analysis methods has been evaluated in an international intercomparison exercise. The focus was on the analysis of microcystins by high-performance liquid chromatography coupled with ultraviolet or photodiode-array detection (HPLC-PDA/UV), currently the most widespread method for microcystin analysis, but the exercise was open for other methods such as enzyme-linked immunosorbent assay (ELISA), protein phosphatase inhibition assay (PPA) and high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS).Thirty-one laboratories from 13 countries participated in the study. For a microcystin-LR (MC-LR) standard solution (S1) of undisclosed quantity, and for a field sample (S3) from a natural cyanobacterial bloom, repeatabilities between 4 and 15% and reproducibilities between 24 and 49% were obtained. No significant differences between single methods were found for S1 and S3, except for a significantly higher repeatability value of ELISA for S1. However, the analysis of microcystins in the field sample (S3) by HPLC-PDA/UV was significantly more variable than for the standard solution (S1). Both the extraction and the analysis of the microcystins appeared to contribute to this variability. It is concluded that standard MC-LR (S1) can be measured with adequate precision by all participating laboratories independently of the method used. With respect to the different methods used the results for the field sample can also be regarded as satisfactory, but clearly showed the need for improvement by standardisation between laboratories. Furthermore, quantification with in-house standards compared to quantification using the supplied MC-LR standard indicated that routine microcystin analysis in laboratories may be also influenced by the variability of available standards, emphasising the need for the production of certified reference materials (CRM).

Chromatography, High Pressure Liquid↗

PubMed2000

Transesophageal 3-dimensional echocardiography: in vivo determination of left ventricular mass in comparison with magnetic resonance imaging.

The objective of this study was to assess the accuracy and reproducibility of transesophageal 3-dimensional echocardiography (3DE) in comparison with magnetic resonance imaging (MRI) for the in vivo calculation of left ventricular mass (LVM). In addition, mass values obtained by M-mode echocardiography were compared with those calculated by MRI. Three-dimensional reconstruction of the left ventricle was performed from a transesophageal and transgastric transducer position with a multiplane transducer in 20 patients. Left ventricular mass was calculated from both transducer positions by using slices of various thicknesses, ranging from 5 to 20 mm. Reproducibility was determined by 5 repeated measurements of mass in each of 5 randomly selected left ventricles. M-mode echocardiography was performed according to the method described by Devereux. For MRI, multiple short-axis views with 10-mm slice thickness were acquired in inspiration hold. Correlation was high for mass determined by 3DE and MRI (for 10-mm slice thickness: r = 0.99; y = 0.99 x - 0.7 g; standard error of estimate = 8.5 g; P <.001). There was no statistical bias, and the limits of agreement ranged from +/-16.4 g to +/-27.2 g, depending on the slice thickness. Variability was lowest for a slice thickness of 10 mm (SD +/- 8.2 g). The reproducibility of mass determination was excellent (mean width of the 95% CI 12.8 g). Left ventricular mass values calculated from the transgastric and transesophageal transducer position were not different from each other (mean bias 0.6 +/- 9.1 g; P = ns). M-mode-based LVM calculations showed systematic overestimation and large measurement variability (bias 23.7 g; 95% CI +/- 92.8 g). Compared with MRI, transesophageal 3DE is an accurate and reproducible method for the determination of LVM and clearly superior to M-mode echocardiography.

Adult↗

PubMed1998

Clinical evaluation of new global clotting assay for monitoring of LMWH treatment: pilot study.

Until now, there has been no reliable method for the monitoring low molecular weight heparin (LMWH) therapy. Based on our observations of the coagulant activity of pseudoserum obtained after clotting of recalcified plasma from patients who were treated with LMWH, we were led to develop a new global clotting assay for the monitoring of this treatment. Methods description. After performing Howell time on PPP, samples were incubated for 30 min in 37 degrees C and they were then centrifuged for 10 min 0.2 ml of this pseudoserum is added to 0.2 ml of citrated normal plasma. Pseudoserum triggered coagulation of normal plasma and the time of new clot formation is expressed in seconds. This assay was called ATHU-test (AHEPA Thromb Haem Unit). We tested 21 normal subjects a in order to define the normal mean value and the range of the method. b) We also checked the reproducibility of the method by repeating the ATHU-test 17 times on the same normal plasma. c) We performed in vitro experiments to study its reliability and we added increasing concentrations using given doses of enoxaparine (4, 8, 12, 16 mg/ml) or fraxiparine (1.5, 3.0, 4.5, 6 u aXa/ml) which was added in vitro to normal plasma confirming the proportional linear regression between duration of our test and the amount of LMWH. Finally d) we checked on the therapy response and the LMWH levels in blood for thrombophiliacs. Results. a) NP, n=21, X=138.6+/-41.1, range 75-225 sec which means that values >X+/-3S=261.9 are distinctively pathological. b) The reproducibility of the method is acceptable, CV=9%. c) It is confirmed that in vitro addition of precautionary doses of LMWH (1.5 u aXa/ml) exceeds the coagulation time of ATHU-test up to 300 sec and it follows a distinctively proportional relationship. d) The monitoring of 20 thrombophiliacs for 2 months proved that: i) All their test times were between 4-15 min. The more we approach the value of 15 min the more danger there is of haemorrhage while, on the contrary, the more values approach 4 min the more the danger of rethrombosis increases. We present the ATHU-test, a simple test which has been used for 6 months now by our Unit, in order to control the LMWH therapy for patients with thromboembolic diseases. The ATHU-test's reproducibility and its small range of normal values, the distinct relationship between therapeutic doses and therapeutic clinical results, as well as the in vitro proof of the linear regression between the coagulation time and the containing amount of LMWH, are likely to establish a new method of choice for the monitoring of LMWH therapy.

Adult↗

PubMed1998

Report of the European DNA Profiling Group (EDNAP)--an investigation of the hypervariable STR loci ACTBP2, APOAI1 and D11S554 and the compound loci D12S391 and D1S1656.

This paper describes the results of three collaborative exercises which continues the EDNAP theme to explore whether uniformity of DNA profiling results could be achieved between European laboratories using STRs. In an earlier exercise, complex hypervariable AAAG-repeat STR loci were investigated, but reproducibility was found to be poor because of the variation of techniques used by participating laboratories. In the exercise reported here, an internal allelic ladder composed of ACTBP2 and D11S554 fragments was distributed. This ladder was used to size ACTBP2 analysed by a "singleplex" PCR amplification and D11S554 combined with APOAI1 in a separate "duplex" reaction. Laboratories were asked to test 7 blood stains, one of which was a known control, and to report the results to the co-ordinating laboratory. The exercise demonstrated that ACTBP2 showed good reproducibility between laboratories, whereas further testing would be needed to validate APOAI1 and D11S554 for interlaboratory comparisons. In separate exercises, the simple loci D12S391 and D1S1656 were tested; both of these showed excellent reproducibility between laboratories.

Alleles↗